Hyaluronic Acid Binding Protein
Bind and detect HA in Multiple Applications
Hyaluronan binding protein (HABP) is a ubiquitously present glycoprotein having specific affinity towards hyaluronic acid (HA). HA, also known as Hyaluronan, is a naturally occurring glycosaminoglycan that is functionally important for the maintenance of a highly hydrated extracellular matrix in tissues, which is involved in cell adhesion and supports cell migration. Hyaluronan also exhibits diverse biological functions by interacting with a large number of hyaluronan-binding proteins and cell surface receptors.
Features
Because of its simplicity and ubiquity, HA is not immunogenic, so classical immunological analysis of HA has not been possible. Instead of hyaluronan/ hyaluronic acid antibodies, a specific and tightly binding protein (Hyaluronic Acid Binding Protein, HA Binding Protein, or HABP) can be used like an antibody to detect or measure HA in multiple applications:
- Histochemistry and Cytochemistry.
- Detection of membrane-transferred Hyaluronic Acid.
- ELISA (HA Quantification Kit available).
- Flow Cytometry.
- HA purification with affinity column.
Download Protocols for biotin HABP from bovine nasal cartilage
Real World Application
Read about the use of Hyaluronic Acid in investigating the cancer resistance of the naked mole-rat in a collaborative effort between the University of Newcastle and the University of Cambridge, UK
Real World Application
In our round up on innovative research, read about Columbia University using AMSBIO's Hyaluronan Binding Protein in their groundbreaking study on engineered wearable edgeless skin constructs (WESCs).
HABP FAQs
- Because of its simplicity and ubiquity, HA is not immunogenic, so classical immunological analysis of HA has not been possible. Instead, a specific and tightly binding protein (Hyaluronic Acid Binding Protein, or HABP) can be used like an antibody to bind or detect HA in multiple applications.
- Most applications require the biotin-HABP, where the VG1 domain binds to HA (hence: HA Binding Protein, HABP for short), then you use labelled streptavidin to get a signal with the biotin as to where/how much of the HABP is there, bound to HA.
- You can’t use a labelled secondary antibody as you might do with a “regular” unconjugated primary antibody, because HABP is not an antibody; and any labelled anti-VG1 antibody would recognize endogenous versican as well as exogenous HABP.
- We recommend unconjugated HABP for HA capture in ELISA-like application. Potentially, it could be used for HA capture in affinity columns – or can also be used if you need to label your HABP with a different tag.
- Hyaluronic Acid has a simple conserved structure, so our HABP has been used to bind or detect HA from a wide variety of species, including: human, mouse, rat, bovine, porcine, canine, naked mole-rat and transgenic Synechococcus sp. cyanobacterium, genetically modified to overexpress Pasteurella multocida HA-synthase (pmHAS) and secrete HA into the culture medium.
- Seyfried et al (2006) paper in Matrix Biology (vol 25: pp 14-19), our HABP (which is Versican G1 domain) can bind Hyaluronan Oligosaccharides or Polysaccharides above minimum size of 10mer – though the three-dimensional HA molecule recognizing ability of G1 domain of HABP is affected not only by the molecular weight but also by the condition of molecules. Smaller molecules may be more affected by their environment.
Versican G1-domain as HA Binding Protein
AMSBIO’s rHABP was launched as a replacement for the HABP formerly available from Seikagaku. It is produced as a recombinant protein in E. coli BL21(DE3)RIL transfected with human versican G1-domain expression vector pRK172VG1 and is available in purified and biotinylated formats.
rHABP specifically detects Hyaluronic Acid (HA), binding to HA from all species and tissues. It does not cross-react to other Glycosaminoglycans or to DNA.
Recombinant production means that our rHABP has higher safety compared with animal-derived HABP.
See Clark et al (2011) details on expression and comparison with Seikagaku's biotin-HABP from bovine cartilage, carried out by Tony Day and colleagues at Manchester. This paper also cites a number of our antibodies to specific glycosaminoglycans: 10E4 (pan Heparan Sulfate), 1B5, 2B6, 3B3 (Chondroitin Sulfate & Dermatan Sulfate 'stubs’), LY111 (native Chondroitin-4-Sulfate) and 5D4 (pan Keratan Sulfate).
The biotin-HABP product supplied by AMSBIO was vital for us to be able to visualise the tissue distribution of hyaluronan in mouse vs. naked mole-rat skin. The images were so brilliant that we have started using biotin-HABP in other studies, e.g. looking at the effects of colitis on colonic hyaluronan distribution.
Citations
The material properties of naked mole-rat hyaluronan.
Kulaberoglu, Y., Bhushan, B., Hadi, F., Chakrabarti, S., Khaled, W., Rankin, K., ... & Frankel, D. (2019). Scientific Reports 9, Article number: 6632.
Perineuronal net formation during the critical period for neuronal maturation in the hypothalamic arcuate nucleus.
Mirzadeh, Z., Alonge, K. M., Cabrales, E., Herranz-Pérez, V., Scarlett, J. M., Brown, J. M., ... & Zeltser, L. M. (2019). Nature Metabolism, 1(2), 212.
Single-nanotube tracking reveals the nanoscale organization of the extracellular space in the live brain.
Godin, A. G., Varela, J. A., Gao, Z., Danné, N., Dupuis, J. P., Lounis, B., ... & Cognet, L. (2017). Nature Nanotechnology 12, 238–243.
Dendritic cells enter lymph vessels by hyaluronan-mediated docking to the endothelial receptor LYVE-1.
Johnson, L. A., Banerji, S., Lawrance, W., Gileadi, U., Prota, G., Holder, K. A., ... & Jackson, D. G. (2017). Nature Immunology.
AAV vector-mediated secretion of chondroitinase provides a sensitive tracer for axonal arborisations.
Alves, J. N., Muir, E. M., Andrews, M. R., Ward, A., Michelmore, N., Dasgupta, D., ... & Rogers, J. H. (2014) Journal of neuroscience methods, 227, 107-120.
Genome-Wide Association Study of Golden Retrievers Identifies Germ-Line Risk Factors Predisposing to Mast Cell Tumours.
Arendt, M. L., Melin, M., Tonomura, N., Koltookian, M., Courtay-Cahen, C., Flindall, N., ... & Murphy, S. (2015). PLoS Genet, 11(11), e1005647.
Molecular composition and expression pattern of the extracellular matrix in a mossy fiber-generating precerebellar nucleus of rat, the prepositus hypoglossi.
Gaál, B., Kecskes, S., Matesz, C., Birinyi, A., Hunyadi, A., & Rácz, É. (2015) Neuroscience letters, 594, 122-126.
Hyaluronan accumulates with high-fat feeding and contributes to insulin resistance.
Kang, L., Lantier, L., Kennedy, A., Bonner, J. S., Mayes, W. H., Bracy, D. P., ... & Wasserman, D. H. (2013). Diabetes, 62(6), 1888-1896
Early Genetic Restoration of Lubricin Expression in Trangenic Mice Mitigates Chondrocyte Peroxynitrite Release and Caspase-3 Activation.
Larson, K. M., Zhang, L., Badger, G. J., & Jay, G. D. (2017). Osteoarthritis and Cartilage.
The oncolytic adenovirus VCN-01 as therapeutic approach against pediatric osteosarcoma.
Martínez-Vélez, N., Xipell, E., Vera, B., Zalacain, M., Marrodan, L., Gonzalez-Huarriz, M., ... & Alonso, M. M. (2015). Clinical Cancer Research, clincanres-1899.
Heterogeneous expression of extracellular matrix molecules in the red nucleus of the rat.
Rácz, É., Gaál, B., & Matesz, C. (2016). Neuroscience , 322, 1-17.
Molecular weight specific impact of soluble and immobilized hyaluronan on CD44 expressing melanoma cells in 3D collagen matrices.
Sapudom, J., Ullm, F., Martin, S., Kalbitzer, L., Naab, J., Möller, S., ... & Pompe, T. (2017). Acta Biomaterialia.
